Driving efficiency in a high-throughput metabolic stability assay through a generic high-resolution accurate mass method and automated data mining

Abstract

Improving analytical throughput is the focus of many quantitative workflows being developed for early drug discovery. For drug candidate screening, it is common practice to use ultra-high performance liquid chromatography (U-HPLC) coupled with triple quadrupole mass spectrometry. This approach certainly results in short analytical run time; however, in assessing the true throughput, all aspects of the workflow needs to be considered, including instrument optimization and the necessity to re-run samples when information is missed. Here we describe a high-throughput metabolic stability assay with a simplified instrument set-up which significantly improves the overall assay efficiency. In addition, as the data is acquired in a non-biased manner, high information content of both the parent compound and metabolites is gathered at the same time to facilitate the decision of which compounds to proceed through the drug discovery pipeline.

An Automated High-Throughput Metabolic Stability Assay Using an Integrated High-Resolution Accurate Mass Method and Automated Data Analysis Software

“…These assays typically use 96-well technologies for incubations and sample preparation and ultraperformance liquid chromatography/mass spectrometry (UPLC/MS) for data acquisition (Korfmacher et al, 1999;Di et al, 2003), which would require more than 50 separate analyses for each enzyme. More recently, high-resolution mass spectrometers have been used for analysis; however, data extraction and data analysis remain cumbersome (O'Connor et al, 2006;Shui et al, 2011). These existing methods, although useful, are semiautomated at best and have their limitations.…”

Exploring skyline for both MSE‐based label‐free proteomics and HRMS quantitation of small molecules

“…High‐resolution MS (HRMS) coupled with LC has been charted as an essential technology for high‐throughput and quantitative analysis of various biological molecules as well as chemical entities in proteomics , metabolomics , and drug metabolism and pharmacokinetic (DMPK) studies . In these applications, MS is used to determine the relative or absolute amounts of analytes among different samples so as to characterize a variety of physiological conditions .…”